3.Construction of siRNA expression vector targeting IGF-1R and its ability to induce cell apoptosls in hmnan lung cancer cells
Ai-Qiang DONG ; Zhi-Yuan MA ; Min-Jian KONG ;
Chinese Journal of Thoracic and Cardiovascular Surgery 2003;0(04):-
Objective Insulin-like growth factor-1 receptor (IGF-1R),similar to insulin receptor,is one of the families of re- ceptor tyrosine kinases.,which has been found to be overexpressed in a variety of cancer.It is the main proliferation and survival sig- nal molecule in cancer cell and plays an important role in cancer growth and progress.Blocking signal transduction of IGF-1R by vari- ous strategies can suppress tumor growth and induce regression of established tumor.This study is to construct the siRNA expression vector targeting IGF-1R and to evaluate its ability to induce cell apoptosis in human lung cancer cells.Methods Two siRNA expres- sion vector,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 targeting IGF-1R,were constructed using pENTR/U6 vector,and a vector targeting hieiferase gene,pENTR/U6-shRNA-Iuc,was constructed as control.After vectors were transfected into A549 for 48h, knockdown of IGF-1R mRNA and protein and Akt phosphorylation were accessed,and DNA ladder and flow cytometry were used for cell apoptosis.Results siRNA expression vectors targeting IGF-1R were successfully constructed,which was confirmed by PCR and DNA sequencing,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 demonstrated the expression were (22.1?2.5) % and (80.1? 3.9) % in IGF-1R mRNA level,(15.2?3.1)% and (47.1?4.1)% in protein level,respectively,compared with pENTR/U6- shRNA-luc.Suppression of IGF-1R by pENTR/U6-shRNA-1 blunted Akt phosphorylation,increased cell apoptosis induced by 3% ethanol,and retained 77.5 % A549 cells in the G0/G1 phase.Conclusion siRNA expression vector targeting IGF-1R can effectively suppress the expression of IGF-1R expression in A549.This study suggests that DNA vector-based RNAi has the potential to be effec- tive and practical cancer gene therapy strategy.
4.Study on Quantitative Diagnosis of Stagnation of Qi and Blood Stasis Syndrome in Chronic Prostatitis
Jia-Chen DONG ; Yi-Ming SUN ; Zhi-Qiang WANG ;
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(03):-
Objective To discuss the quantitative diagnosis of stagnation of Qi and blood stasis syndrome in chronic prostatitis. To make diagnosis chart and ensure diagnosis threshold level which provide statistics evidence for syndrome differentiation of TCM. Methods By the statistical ways, 168 cases of chronic prostatitis belong to stagnation of Qi blood and stasis syndrome and 198 cases of non-stagnation of Qi and blood stasis syndrome were investigated. To make a diagnosis chart and ensure diagnosis threshold level by applying the method of the maximum likelihood discriminatory analysis. Results The quantitative diagnosis chart was made and diagnosis threshold level was 26. According to the retrospective and prospective test, its sensitivity, especially degree, coincidence rate, error rate and positive likelihood ratio were 94.64%, 88.89%, 91.53%, 8.47%, 8.52 and 94.28%, 90.32%, 92.42%, 7.58%, 9.74. Conclusion The indexes of the quantitative diagnosis have good objectivity. According to the retrospective and prospective test, the diagnosis chart was proved to be practical.
5.A pilot study on purification of human plasminogen from cord blood plasma and inducing posterior vitreous detachment
Wei-zhi, CHEN ; Jian-qiang, DONG ; Xiao-xin, LI
Chinese Journal of Experimental Ophthalmology 2011;29(8):690-696
Background Many studies and clinical trials of pharmacologic vitreolysis are already under way to try to improve vitreo-retinal surgery and to liquefy and detach the vitreous from the retina ultimately, including chondroitinase,hyaluronidase,dispase and plasmin. However, there has not been any report on purification of human plasminogen from cord blood plasma and inducing posterior vitreous detachment of the animal eye at present.Objective This study was designed to isolate and purify the production of human plasminogen (Plg) from cord blood plasma with ethanol precipitation and evaluate the efficacy of Plg in inducing posterior vitreous detachment (PVD).Methods Human Plg was Separated and purified from cord blood plasma by ethanol precipitation method. The protein band corresponding to Plg with molecular mass of 92 000 was revealed in SDS-PAGE and confirmed by MALDI-TOF and Mascot database. Anion-exchange chromatography and plasminogen activity assay kit were used to obtain purified Plg with biological activity. Twenty-five fresh pig eyes were enucleated and assigned to 5 groups and 5 eyes for each group. The normal eyes were used as control group. Balanced salt solution(BSS)of 0.1 ml was intravitreally group and standard substance group. All of the eyes were then incubatedfor 60 minutes under the 37 ℃. Retinal histopathology and ultrastructure were examined under the light microscopy, scanning electron microscopy ( SEM ) and transmission electron microscopy (TEM). Results The Plg with potential fibrinolytic activity was successfully extracted and purified from cord blood plasma by ethanol precipitation method. No posterior vitreous detachment (PVD) was seen in normal control group, BSS group and r-SK group following the intravitreal injection under the sem. However,PVD was demonstrated in r-SK+ Plg group and standard substance group under the SEM. The inner limiting membrane ( ILM ) and the retina were well preserved in all of the experimental eyes. No retinal morphology and ultrastructural abnormality were found under the light and SEM and TEM. Conclusion Ethanol precipitation is a feasible way to isolate and purify Plg from human cord blood plasma. Extracted Plg shows potential fibrinolytic intravitreal injection of Plg.
6.Meta-analysis of internal fixation versus hip replacement in the treatment of trochanteric fractures.
Jian-Bin DONG ; Zhi-Yong WANG ; Hao LU ; Yuan TIAN ; Xin-Rui WANG ; Zhi-Qiang ZHANG
China Journal of Orthopaedics and Traumatology 2015;28(3):245-251
OBJECTIVETo compare the efficacy of internal fixation (including PFNA and PFN) versus hip replacement (including FHR or THA) in the treatment of trochanteric fractures in adults.
METHODSReports of studies using randomized controlled trials (RCT) to compare internal fixationg with hip replacement in the management of intertrochanteric fractures were retrieved (up to January 1, 2013) from the Cochrane Library, PUBMED Data, CNKI (China National Knowledge infrastructure), Elsevier, the Chinese Biomedical Database, Wanfang Data, and manually. Methodological quality of the trials was critically assessed, and relevant data were extracted. Statistical software RevMan 5.0 was used for data-analysis.
RESULTSSeven articles were included in the meta-analysis. The results showed that,compared internal fixation with hip replacement,there were statistical significance in the duration of surgery time [WMD = -2.66, 95% CI (-5.25,-0.06), P = 0.05], intra-operative blood loss [WMD = -24.20, 95% CI (-30.38, -18.02), P < 0.000 01], hospital stays time [WMD = -4.72, 95% CI (-5.18, -4.25), P < 0.000 01], bearing load time [WMD = -29.54, 95% CI (-30.77, -28.31), P < 0.000 01], total complications rate [WMD = 0.15, 95% CI (0.11, 0.22), P < 0.000 01], the good rate of Harris scores [WMD = 1.09, 95% CI (0.54,1.32), P < 0.05]. However, there were no statistical significance in the rate of deep venous thrombosis [WMD = 1.09, 95% CI (0.47, 2.55), P > 0.05]. CON- CLUSION: Hip replacement (containing FHR or THA) for the treatment of intertrochanteric fractures is superior to internal fixa- tion in regards to the duration of surgery time, the mean duration of hosipital stays, mean post-operative down time, intra-opera- tive blood loss, the rate of post-operative good Harris scores. But there is not enough evidence to show any difference between hip replacement (containing THA or FHR) and internal fixation in regards to the rate of deep venous thrombosis. However, internal fixation for the treatment of intertrochanteric fractures is superior to hip replacement (containing FHR or THA) in regards to total complications rate.
Arthroplasty, Replacement, Hip ; methods ; Fracture Fixation, Internal ; methods ; Hip Fractures ; surgery ; Humans
7.Using the SELDI Protein Chip System to Detect Changes in Protein Expression in Vero Cells after Infection
Zhi-jun, LIU ; Bin, WANG ; Zhi-yong, YAN ; Xu-xia, SONG ; Dong-meng, QIAN ; Zhi-qiang, BAI
Virologica Sinica 2007;22(1):68-73
Human herpes simplex virus 1 (HSV-1) causes facial,ocular,and encephalitic disease and is associated with latent infection and cancer.Here,we developed a means of studying the pathogenesis of HSV-1 infection at the protein level by using the SELDI Protein Chip to detect changes of protein expression in Vero cells cultured in vitro.After infection with HSV-1 and culture for 12,24 or 48 h,cells were harvested and lysed.IMAC3 arrays were applied to SELDI-TOF-MS to detect proteomic differences before and after infection.The chip detected a series of differentially expressed protein peaks.Interestingly,both peaks at 16 912 Da and 17 581 Da corresponded precisely with the molecular mass of ISG 15,which may participate in antiviral activity during the process of infection.Thus,the results we obtained can serve as a basis to study the pathogenesis of HSV-1 and the interaction between the virus and its host.In addition,they can help in the discovery of new therapeutic targets for treatment of HSV-1 infection.
9.Effects of simvastatin on the expression of RANTES in patients with hypercholesterolemia
Yong-Hong LI ; Zhi-Ming GE ; Zhi-Qiang LI ; Shan-Lang CAI ; Yi AN ; Qi-Xin WANG ; Guo-Xiong DONG ;
Chinese Journal of Emergency Medicine 2006;0(12):-
6.24 mmol/L) and sixty healthy persons in the health center of our hospital were investigated as hyperhpidemia group (Hyperlipidemias) and control group (Controls) respectively.Hyperlipidemias were given simvastatin 20 mg?d~(-1) for twelve weeks (Statins).Blood samples of ulnar vein were extracted from Statins at the end of twelve weeks as well as Controls and Hyperhpidemias at the beginning of the experiment. Blood serum,plasma and mononuclearcell were extracted and stored at a refrigerator of-80℃.The level of plasma angiotensinⅡwas detected by the method of radioimmunity.While the expression of RANTES mRNA and protein on mononuclearcell were assessed by real time reverse transcription polymerse chain reaction and Western blot respectively.Results①The plasma angiotensinⅡof Hyperlipidemias was higher than that of Controls [(92.13?22.03) vs (50.85?12.12),P
10.Applied research of intermaxillary fixation screw in the jaw fracture.
Wei HUANG ; Zhi-qiang CAO ; Dong FANG ; Zhen-yu HU
Chinese Journal of Plastic Surgery 2004;20(5):364-365
OBJECTIVETo evaluate and generalize the application of intermaxillary fixation screw in the jaw fracture.
METHODS41 cases of jaw fracture have been treated with intermaxillary fixation screw.
RESULTSBoth function and appearance have recuperated in 40 cases except 1 case has light malocclusion.
CONCLUSIONSApplication of intermaxillary fixation screw advanced the traditional therapy.
Adolescent ; Adult ; Bone Screws ; standards ; Female ; Humans ; Jaw Fractures ; surgery ; Male ; Maxillary Fractures ; surgery ; Middle Aged ; Orthopedic Fixation Devices ; standards ; Reconstructive Surgical Procedures ; instrumentation ; methods ; Treatment Outcome